Isotope Substitution of Promiscuous Alcohol Dehydrogenase Reveals the Origin of Substrate Preference in the Transition State.
Isotope Substitution of Promiscuous Alcohol Dehydrogenase Reveals the Origin of Substrate Preference in the Transition State.
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DOI：
10.1002/anie.201712826
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发表时间：
2018-03-12
期刊：
影响因子：
--
通讯作者：
Allemann RK
中科院分区：
文献类型：
--
作者：
Behiry EM;Ruiz-Pernia JJ;Luk L;Tuñón I;Moliner V;Allemann RK
通过对嗜热脂肪芽孢杆菌 (BsADH) 的醇脱氢酶进行酶同位素标记，研究了混杂酶中底物偏好的起源。在生理温度下，蛋白质与反应坐标的动态耦合不显着。然而，动态耦合的程度在较低温度下高度依赖于底物。对于苯甲醇，观察到大于 1 的酶同位素效应，而对于异丙醇，酶同位素效应接近 1。对过渡态的频率运动分析表明，在催化空间大的醇期间，活性位点周围的残基经历了显着的位移。 BsADH 更喜欢较小的底物，这会导致沿反应坐标的蛋白质摩擦较小，并降低动态重交叉的频率。该假设可以预测酶同位素对多种底物的影响趋势。
The origin of substrate preference in promiscuous enzymes was investigated by enzyme isotope labelling of the alcohol dehydrogenase from Geobacillus stearothermophilus (BsADH). At physiological temperature, protein dynamic coupling to the reaction coordinate was insignificant. However, the extent of dynamic coupling was highly substrate‐dependent at lower temperatures. For benzyl alcohol, an enzyme isotope effect larger than unity was observed, whereas the enzyme isotope effect was close to unity for isopropanol. Frequency motion analysis on the transition states revealed that residues surrounding the active site undergo substantial displacement during catalysis for sterically bulky alcohols. BsADH prefers smaller substrates, which cause less protein friction along the reaction coordinate and reduced frequencies of dynamic recrossing. This hypothesis allows a prediction of the trend of enzyme isotope effects for a wide variety of substrates.
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影响因子：
15
作者：
Luk, Louis Y. P.;Loveridge, E. Joel;Allemann, Rudolf K.
通讯作者：
Allemann, Rudolf K.
影响因子：
4.8
作者：
Nagel, Zachary D.;Cun, Shujian;Klinman, Judith P.
通讯作者：
Klinman, Judith P.
影响因子：
15
作者：
Ruiz-Pernia JJ;Luk LY;García-Meseguer R;Martí S;Loveridge EJ;Tuñón I;Moliner V;Allemann RK
通讯作者：
Allemann RK
影响因子：
2.9
作者：
Ceccarelli, C;Liang, ZX;Bahnson, BJ
通讯作者：
Bahnson, BJ
DOI：
10.1073/pnas.1111252108
发表时间：
2011-08-23
影响因子：
11.1
作者：
Adamczyk, Andrew J.;Cao, Jie;Warshel, Arieh
通讯作者：
Warshel, Arieh